CN110317754A - A kind of microbial liquid microbial inoculum and preparation method thereof containing polyglutamic acid - Google Patents
A kind of microbial liquid microbial inoculum and preparation method thereof containing polyglutamic acid Download PDFInfo
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- CN110317754A CN110317754A CN201910575295.8A CN201910575295A CN110317754A CN 110317754 A CN110317754 A CN 110317754A CN 201910575295 A CN201910575295 A CN 201910575295A CN 110317754 A CN110317754 A CN 110317754A
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- China
- Prior art keywords
- polyglutamic acid
- microbial
- microbial inoculum
- liquid
- chlorophyll
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- 108010020346 Polyglutamic Acid Proteins 0.000 title claims abstract description 50
- 229920002643 polyglutamic acid Polymers 0.000 title claims abstract description 48
- 239000002068 microbial inoculum Substances 0.000 title claims abstract description 45
- 230000000813 microbial effect Effects 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
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- 238000000576 coating method Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
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- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 229910000388 diammonium phosphate Inorganic materials 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
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- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
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- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
- A01K63/04—Arrangements for treating water specially adapted to receptacles for live fish
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
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- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Environmental Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Husbandry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of microbial liquid microbial inoculum and preparation method thereof containing polyglutamic acid, including 1-20 parts of Promoting bacteria bacterium solution, 1-20 parts of polyglutamic acid, 1-20 parts of minerals, 50-70 parts of biological organic matter, 1-5 parts of chlorophyll, 1-5 parts of vitamin.Complex microorganism liquid bacterial agent of the invention, water-soluble speed is fast, and viable bacteria content is high, can inhibit pathogenic microorganism；The harmful substance that can be flocculated in water body, purifies water；The heavy metal ion in water body can be chelated, heavy metal ion toxicity is alleviated；It can promote the photosynthesis of algae, provide nutrition abundant for aquatic animal, there are the multi-efficiencies such as water purification, volume increase.
Description
Technical field
The present invention relates to microbial bacterial agent technical fields, and in particular to a kind of micro- life suitable for aquaculture containing polyglutamic acid
Object liquid bacterial agent and preparation method thereof.
Background technique
Intensive aquaculture at home and abroad rapidly develops in recent years, and a large amount of bait are in the form of Metabolism of Fishes product and surplus bait
Into environment, the pollution of water body, sediment of pond etc. is caused.Cultivated water ecological functions are degenerated, and disease is on the rise, and are supported to aquatic products
It grows industry and causes huge economic losses.
The common method of water pollution processing includes method of chemical treatment, physical treatment process and biological treatment, wherein is used
Biological treatment processing is increasingly extensive.However, existing most of biological treatment validity of products for being used to handle water pollution
Low, water quality that treated is not still up to standard, or even the phenomenon that aquatic products mortality occurs, causes to aquaculturist more serious
Economic loss.
Summary of the invention
Goal of the invention: in order to solve, the existing effective bacterial content of microbial liquid microbial inoculum is low, easy in inactivation and production in transportational process
The problem of product validity difference, the present invention provide a kind of microorganism suitable for aquaculture, with multi-efficiencies such as water purification, volume increase
Liquid bacterial agent.
In order to achieve the above object, the invention discloses a kind of microbial liquid microbial inoculum containing polyglutamic acid, including it is as follows
Component by weight: 1-20 parts of Promoting bacteria bacterium solution, 1-20 parts of polyglutamic acid, 1-20 parts of minerals, biological organic matter 50-70
Part, 1-5 parts of chlorophyll, 1-5 parts of vitamin.
Specifically, in above-mentioned technical proposal, the Promoting bacteria bacterium solution is bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens any one fermentation liquid or two kinds of fermentations) and in bacillus subtilis (Bacillus subtilis)
The combination of liquid, the viable count in the bacillus amyloliquefaciens fermentation liquid or bacillus subtilis fermentation liquor are 1 × 109-1×
1011CFU/mL.The additive amount of Promoting bacteria bacterium solution is according to any one in bacillus amyloliquefaciens and bacillus subtilis or combination
Summation calculate, viable count >=6 × 10 of the Promoting bacteria bacterium solution in microbial liquid microbial inoculum9CFU/mL。
The bacillus amyloliquefaciens (Bacillus amyloliquefaciens) are in China typical culture collection
The deposit number of the heart is CCTCC NO:M 2016266, and bacillus subtilis (Bacillus subtilis) is trained in Chinese Typical Representative
The deposit number for supporting object collection is CCTCC NO:M 2016264, is recorded in patent 2016106658324.
Further, the Promoting bacteria bacterium solution be bacillus amyloliquefaciens fermentation liquid and bacillus subtilis fermentation liquor according to
Volume ratio (0.1-20): the mixed liquor of (0.1-20) mixing.
Specifically, the polyglutamic acid weight average molecular weight is 4-3000kDa, preferred molecular weight 4-60kDa.The poly- paddy
Propylhomoserin be polyglutamic acid fermentation liquid, polyglutamate sterling, polyglutamic acid wettable powder, polyglutamic acid fermentation liquid dried object and
Any one or the combination of several of them in polyglutamic acid sterling.The additive amount of polyglutamic acid is according to polyglutamic acid fermentation liquid, poly- paddy
Any one in propylhomoserin salt sterling, polyglutamic acid wettable powder, polyglutamic acid fermentation liquid dried object and polyglutamic acid sterling
Or polyglutamic acid gross mass contained in combination calculates.
Specifically, the minerals are potassium, iron, copper, zinc, manganese, nickel, molybdenum, cobalt, tin, magnesium, sodium, calcium, chlorine, iodine, sulphur, oxygen, phosphorus
With the pure compounds of any two kinds of element composition described above in selenium or the mixture of two or more pure compounds.Mineral
The additive amount of matter is calculated according to minerals gross mass contained in the mixture of pure compounds or pure compounds.
Specifically, the biological organic matter is the combination of any one in molasses extract and yeast extract or two kinds.
The additive amount of biological organic matter is calculated according to any one in molasses extract and yeast extract or combined gross mass.
Specifically, the chlorophyll is chlorophyll a, chlorophyll b, Chlorofucsin, CHLOROPHYLL d, chlorophyll f, bacteriochlorophyll
With any one or the combination of several of them in protochlorophyll.The additive amount of chlorophyll according to chlorophyll a, chlorophyll b, Chlorofucsin,
Any one in CHLOROPHYLL d, chlorophyll f, bacteriochlorophyll and protochlorophyll or combined gross mass calculating.Chlorophyll is to plant
Object carries out photosynthetic primary pigments, is located at thylakoid membrane, plays the role of a nucleus in photosynthetic light absorption, have simultaneously
It is provided with the multiple uses such as vitamin, hematopoiesis, disease-resistant, removing toxic substances.
Specifically, the vitamin is vitamin A, vitamin B, vitamin C, vitamin D, biotin, vitamine M, dimension
Any one or the combination of several of them in raw element P.The additive amount of vitamin is given birth to according to vitamin A, vitamin B, vitamin C, dimension
Plain D, biotin, vitamine M, any one or combined gross mass in citrin calculate.Vitamin is that aquaculture is dynamic
Object maintains trace organic substance necessary to normal physiological function, neither offer energy, is also not involved in composition cell, but in water
It produces and plays an important role in cultivated animals growth, metabolism and growth course.
Further, the present invention also provides the preparation methods of mentioned microorganism liquid bacterial agent, specifically includes the following steps:
(1) Promoting bacteria that inclined-plane saves is inoculated on activation medium, 25-35 DEG C of activation 18-24h, picking activation training
The strain on base is supported, is transferred in fermentation medium, fermentation liquid or fermentation concentrated liquor are placed in 4 by 30-35 DEG C of culture 12-24h
Low-temperature treatment 28-50h in DEG C freezer, obtains Promoting bacteria bacterium solution.When Promoting bacteria includes two kinds, then fermentation liquid or hair are respectively obtained
Then zymotic fluid concentrate mixes fermentation liquid according to volume ratio.The activation medium is LB culture medium, is formulated as follows:
Peptone 10g/L, yeast powder 5g/L, sodium chloride 10g/L, pH 6.0-8.0, solvent is water.The formula of the fermentation medium is such as
Under: carbon source 10-100g/L, nitrogen source 1-40g/L, inorganic salts 0.01-50g/L, pH 5.0-9.0, solvent is water.The fermentation training
The carbon source for supporting base is to appoint in glucose, sucrose, maltose, lactose, xylose, fructose, lactic acid, citric acid, glycerol, starch and molasses
The combination for one or more of anticipating；The nitrogen source is yeast powder, beef extract, peptone, yeast extract, corn pulp, beancake powder, cottonseed cake
Powder, urea, (NH4)2SO4、NH4Cl、(NH4)2HPO4And NH4NO3Middle any one or the combination of several of them；The inorganic salts are sulphur
Any one or the combination of several of them in hydrochlorate, phosphate, dihydric phosphate, phosphoric acid hydrogen disalt and hydrochloride.
(2) the Promoting bacteria bacterium solution for obtaining step (1) and polyglutamic acid, minerals, biological organic matter, chlorophyll and Wei Sheng
Element is according to mass parts than mixing to get microbial liquid microbial inoculum.
The beneficial effect comprise that
1, the Promoting bacteria bacterium solution in microbial liquid microbial inoculum can effectively inhibit Aeromonas sobria in water body, thermophilic aqueous vapor list
The growth of other pathogenic bacteria such as born of the same parents bacterium, Aeromonas caviae, Aeromonas veronii and tarda, effective prevention and control are by pathogen
The aquiculture animal disease of initiation, especially bacillary hemorrhage, to protect aquatic animal healthy growth.
2, polyglutamic acid is generated by microbial fermentation, passes through the homopolymerization of glutamy amine key by D-Glu and Pidolidone monomer
It forms, there is nontoxic, biodegradable and advantages of environment protection, provide pod membrane for the Promoting bacteria in microbial bacterial agent and protect
Shield makes Promoting bacteria form biofilm system, reaches a kind of stable equilibrium state, be at gemma state；Polyglutamic acid rich in-
The various actives group such as COOH ,-CO- ,-NH-, charge density is high, and molecular weight is larger and linear, and there is good flocculation to live
Property, the harmful substance in the water body that can flocculate purifies water, and can chelate the heavy metal ion in water body, alleviates heavy metal ion
Toxicity.
3, minerals are applied in water body, can promote the photosynthesis of algae, and nutrient growth and the reproduction for improving algae are raw
Long, it is to increase primary productivity in itself, in addition, it can guarantee algae too that the mechanism of action, which is different from plant hormone,
Part photosynthesis is carried out in the environment such as sunlight line is weak, water body is muddy, night, effect is better than other similar products.
4, biological organic matter inhibits the sprouting of Promoting bacteria gemma by providing high organic environment and hyperosmosis environment, from
And Promoting bacteria is made to be in gemma state, and when microbial liquid microbial inoculum is diluted application, organic matter can be Promoting bacteria bud again
The sprouting of spore provides nutrition, promotes the fast breeding of Promoting bacteria.When biological organic matter and polyglutamic acid are used in combination, except having
Outside nutrition synergistic function, the main vigor that Promoting bacteria in liquid bacterial agent is maintained by synergistic effect substantially prolongs microorganism liquid
The shelf life of body microbial inoculum, can effectively solve that the existing effective bacterial content of microbial liquid microbial inoculum is low, easy in inactivation and production in transportational process
The problem of product validity difference.
Detailed description of the invention
The present invention is done with reference to the accompanying drawings and detailed description and is further illustrated, of the invention is above-mentioned
And/or otherwise advantage will become apparent.
Fig. 1 is the number of viable survival curve of different microbial inoculums in embodiment 3.
Specific embodiment
According to following embodiments, the present invention may be better understood.
In following embodiment, bacillus amyloliquefaciens (Bacillus amyloliquefaciens) are in Chinese Typical Representative culture
The deposit number of object collection is CCTCC NO:M 2016266.
Bacillus subtilis (Bacillus subtilis) is in the deposit number of China typical culture collection center
CCTCC NO:M 2016264 is recorded in patent 2016106658324.
The Related product of polyglutamic acid is prepared by existing fermentation technique.
Minerals, biological organic matter, chlorophyll, vitamin are bought from market or according to prior art preparations.
The fermentation of 1 Promoting bacteria of embodiment
Promoting bacteria bacterium solution is prepared as follows:
Bacillus amyloliquefaciens CCTCC NO:M 2016266, the bacillus subtilis CCTCC NO that 4 DEG C of inclined-planes are saved:
M 2016264 is activated, and is inoculated on eggplant shape culture bottle culture medium (i.e. activation medium) respectively, 32 DEG C of culture 18h.By activation
Strain is transferred in fermentation medium respectively, and fermentation liquid for 24 hours, is placed in low-temperature treatment 48h in Cool Room 4 DEG C by 30 DEG C of cultures, through flat
Plate coating measures, and the number of viable in every kind of microbial fermentation solution can reach 8 × 109CFU/mL or more sends out two kinds of microorganisms
Zymotic fluid is compounded according to the ratio mixing of volume ratio 1:1 up to Promoting bacteria bacterium solution.
Culture medium in the eggplant shape culture bottle is LB culture medium, fermentation medium component are as follows: glucose 50g/L, albumen
Peptone 10g/L, (NH4)2SO40.05g/L, NaCl 1g/L, MnSO4·4H2O 0.5g/L, KH2PO41.0g/L, pH 6.0-
8.0。
The ventilatory capacity of filtrated air is 0.6vvm, mixing speed 220rpm during the fermented and cultured of the above production tank.
The preparation of 2 complex microorganism liquid bacterial agent of embodiment
It is prepared as follows bacterium solution containing Promoting bacteria, polyglutamic acid, minerals, biological organic matter, chlorophyll, vitamin
Complex microorganism liquid bacterial agent:
(1) polyglutamic acid prepares (bibliography Xu Z, Feng X, Dan Z, et al.Enhanced using fermentation method
poly(γ-glutamic acid)fermentation by Bacillus subtilis,NX-2immobilized in an
Aerobic plant fibrous-bed bioreactor [J] .Bioresour Technol, 2014,155:8-14), fermentation
Polyglutamic acid content is 15% after liquid is concentrated；
(2) minerals are by market purchasing；
(3) molasses are purchased by sugar refinery；
(4) chlorophyll is by market purchasing；
(5) vitamin is by market purchasing；
(6) the compound Promoting bacteria bacterium solution for obtaining embodiment 1 and polyglutamic acid Fermented Condensed liquid, minerals, molasses, Ye Lv
Element, vitamin are uniformly mixed according to the ratio that mass ratio is 10:5:1:60:1:1 obtains complex microorganism liquid bacterial agent.Through examining
It surveys, polyglutamic acid content mass percent is 3.2%, and minerals mass percent is 1.5%, content of organic matter mass percent
It is 65%, chlorophyll mass percent is 1.5%, and vitamin mass percent is 1%, and living bacteria count is 7.6 × 109CFU/
mL。
3 complex microorganism liquid bacterial agent of embodiment can open preservation up to 18 months
Microbial inoculum used in the present embodiment is as follows:
(1) mix bacterium agent that embodiment 1 is prepared is diluted to living bacteria count is 7.6 × 109CFU/mL, as bacterium
Agent 1.
(2) mix bacterium agent that embodiment 1 is prepared is mixed in proportion with minerals, molasses, chlorophyll, vitamin
It is even, make content of mineral substances 1.5%, the content of organic matter 65%, chlorophyll content 1.5%, vitamin content 1% has
Imitating viable count is 7.6 × 109CFU/mL, as microbial inoculum 2.
(3) mix bacterium agent that embodiment 1 is prepared is mixed in proportion with polyglutamic acid, molasses, chlorophyll, vitamin
Uniformly, make polyglutamic acid content 3.2%, the content of organic matter 65%, chlorophyll content 1.5%, vitamin content is
1%, living bacteria count is 7.6 × 109CFU/mL, as microbial inoculum 3.
(4) mix bacterium agent that embodiment 1 is prepared is mixed in proportion with polyglutamic acid, minerals, chlorophyll, vitamin
It closes uniformly, makes polyglutamic acid content 3.2%, content of mineral substances 1.5%, chlorophyll content 1.5%, vitamin content is
1%, living bacteria count is 7.6 × 109CFU/mL, as microbial inoculum 4.
(5) mix bacterium agent that embodiment 1 is prepared is mixed in proportion with polyglutamic acid, minerals, molasses, vitamin
Uniformly, make polyglutamic acid content 3.2%, content of mineral substances 1.5%, the content of organic matter 65%, vitamin content is
1%, living bacteria count is 7.6 × 109CFU/mL, as microbial inoculum 5.
(6) mix bacterium agent that embodiment 1 is prepared is mixed in proportion with polyglutamic acid, minerals, molasses, chlorophyll
Uniformly, make polyglutamic acid content 3.2%, content of mineral substances 1.5%, the content of organic matter 65%, chlorophyll content is
1.5%, living bacteria count is 7.6 × 109CFU/mL, as microbial inoculum 6.
(7) contain polyglutamic acid, minerals, biological organic matter, chlorophyll, vitamin and Promoting bacteria for what embodiment 2 obtained
Complex microorganism liquid bacterial agent, as microbial inoculum 7.
Above-mentioned 7 kinds of microbial inoculums are placed in 25 DEG C, is stored in the environment of relative humidity 40%, 1mL microbial inoculum was respectively taken every 2 months,
Dilution 108Times, it in applying solid LB culture medium, is placed under 32 DEG C of environment, cultivates 18-24h, obtained using colony counting method effective
Number of viable, continuous sampling 20 months.If three groups of parallel laboratory tests, living bacteria count is changed over time such as Fig. 1 institute in different microbial inoculums
Show.
As shown in Figure 1, the living bacteria count amount of microbial inoculum 1, microbial inoculum 2 and microbial inoculum 4 declines in 20 months rapidly, at the 18th
When the moon, living bacteria count amount is below 2 × 109CFU/mL, the living bacteria count amount of microbial inoculum 3, microbial inoculum 5 and microbial inoculum 6 is the 18th
5 × 10 are still higher than at a month9CFU/mL, but it is lower than the living bacteria count amount of microbial inoculum 7.It can be seen that composite microbial of the invention
The object liquid bacterial agent holding time is longer.
Fish crop can be improved in 4 complex microorganism liquid bacterial agent of embodiment
In the present embodiment containing the compound of polyglutamic acid, minerals, biological organic matter, chlorophyll, vitamin and Promoting bacteria
Microbial liquid microbial inoculum is prepared with embodiment 2.Using orbfish and Tilapia mossambica as subjects, test sets 2 processing, each processing altogether
3 repetitions.The 1 common microbial inoculum (2kg/667m of addition of processing2), 2 addition complex microorganism liquid bacterial agent (2kg/667m of processing2), separately
If not adding control, daily management is all the same, the results are shown in Table 1.
Influence of the 1 complex microorganism liquid bacterial agent of table to orbfish and Rofe fish crop
As shown in Table 1, the yield of orbfish and Tilapia mossambica can be improved by applying common microbial inoculum and complex microorganism liquid bacterial agent,
But complex microorganism liquid bacterial agent becomes apparent from terms of promoting yield.
5 complex microorganism liquid bacterial agent of embodiment can improve meat quality of fish
Measurement item main body is the final fish of embodiment 4 in the present embodiment.As shown in Table 2, common microbial inoculum and compound is applied
Microbial liquid microbial inoculum can improve the total amino acid content and essential amino acids content of orbfish and Tilapia mossambica, be especially to human body
The growth and development of children is of great significance, but complex microorganism liquid bacterial agent promotes effect and becomes apparent from.Meanwhile applying common bacterium
Agent and complex microorganism liquid bacterial agent can reduce the moisture content of orbfish and Tilapia mossambica, and complex microorganism liquid bacterial agent reduces water
Point effect becomes apparent from, this illustrates to add complex microorganism liquid bacterial agent while improving fish crop, and meat quality of fish also can obviously change
It is kind.
Influence of the 2 complex microorganism liquid bacterial agent of table to orbfish and meat quality of java tilapia
The thinking and method of the microbial liquid microbial inoculum that the present invention provides a kind of containing polyglutamic acid and preparation method thereof, tool
Body realizes that there are many method of the technical solution and approach, the above is only a preferred embodiment of the present invention, it is noted that right
For those skilled in the art, without departing from the principle of the present invention, several improvement can also be made
And retouching, these modifications and embellishments should also be considered as the scope of protection of the present invention.Each component part being not known in the present embodiment is equal
It can be realized with the prior art.
Claims (9)
1. a kind of microbial liquid microbial inoculum containing polyglutamic acid, which is characterized in that including following components by weight: growth-promoting
1-20 parts of bacterium bacterium solution, 1-20 parts of polyglutamic acid, 1-20 parts of minerals, 50-70 parts of biological organic matter, 1-5 parts of chlorophyll, vitamin
1-5 parts.
2. microbial liquid microbial inoculum according to claim 1, which is characterized in that the Promoting bacteria bacterium solution is solution starch gemma
The fermentation liquid of bacillus and/or bacillus subtilis；The bacillus amyloliquefaciens fermentation liquid or bacillus subtilis fermentation liquor
Middle viable count is 1 × 109-1×1011CFU/mL；Viable count >=6 of the Promoting bacteria bacterium solution in microbial liquid microbial inoculum ×
109CFU/mL。
3. microbial liquid microbial inoculum according to claim 2, which is characterized in that the Promoting bacteria bacterium solution is solution starch gemma
Bacillus fermentation liquid and bacillus subtilis fermentation liquor are according to volume ratio (0.1-20): the mixed liquor of (0.1-20) mixing.
4. microbial liquid microbial inoculum according to claim 1, which is characterized in that the polyglutamic acid weight average molecular weight is 4-
3000kDa；Polyglutamic acid is polyglutamic acid fermentation liquid, polyglutamate sterling, polyglutamic acid wettable powder, polyglutamic acid hair
Any one or the combination of several of them in zymotic fluid dried object and polyglutamic acid sterling.
5. microbial liquid microbial inoculum according to claim 1, which is characterized in that the minerals be potassium, iron, copper, zinc,
The compound of any two kinds of element composition described above in manganese, nickel, molybdenum, cobalt, tin, magnesium, sodium, calcium, chlorine, iodine, sulphur, oxygen, phosphorus and selenium is pure
The mixture of product or two or more pure compounds.
6. microbial liquid microbial inoculum according to claim 1, which is characterized in that the biological organic matter is molasses extract
With any one in yeast extract or two kinds of combination.
7. microbial liquid microbial inoculum according to claim 1, which is characterized in that the chlorophyll is chlorophyll a, chlorophyll
B, any one or the combination of several of them in Chlorofucsin, CHLOROPHYLL d, chlorophyll f, bacteriochlorophyll and protochlorophyll.
8. microbial liquid microbial inoculum according to claim 1, which is characterized in that the vitamin is vitamin A, vitamin
B, vitamin C, vitamin D, biotin, vitamine M, any one or the combination of several of them in citrin.
9. the preparation method of microbial liquid microbial inoculum described in claim 1, which comprises the following steps:
(1) Promoting bacteria that inclined-plane saves is inoculated on activation medium, 25-35 DEG C of activation 18-24h；Picking activation medium
On strain, transfer in fermentation medium, 30-35 DEG C of culture 12-24h, by fermentation liquid or ferment concentrated liquor be placed in 4 DEG C it is cold
Low-temperature treatment 28-50h in library obtains Promoting bacteria bacterium solution；
(2) the Promoting bacteria bacterium solution for obtaining step (1) and polyglutamic acid, minerals, biological organic matter, chlorophyll and vitamin are mixed
It is even to get.
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